FIG. 6.

Pretreatment of cells with sodium orthovanadate reverses the effect of leishmania infection on c-FOS protein expression. Cells were either untreated or treated with orthovanadate (20 μM) for 3 h and then incubated with leishmania amastigotes at an approximate parasite-to-cell ratio of 15:1. After overnight incubation, control and infected cells were stimulated with PMA (100 nM) for 4 h. Cells were lysed in modified RIPA buffer as described in Materials and Methods. Whole-cell lysates were separated on SDS–7.5% polyacrylamide gels, transferred to nitrocellulose membranes, and probed with c-FOS antibodies. The blots were developed by ECL, and an autoluminogram of a blot is shown. The blot shown is from one of two independent experiments that yielded similar results, and the histogram shows the average values of the two experiments.